Structural studies on dynein-microtubule complex

动力蛋白-微管复合物的结构研究

• 批准号: 7033453
• 负责人: Masahide Kikkawa
• 金额: $ 23.55万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7033453
• 关键词: Chlamydomonas; bioenergetics; bioimaging /biomedical imaging; biophysics; calcium; cilium /flagellum motility; conformation; cryoelectron microscopy; dynein ATPase; gene mutation; high throughput technology; microtubules; nucleotides; plant genetics; protein protein interaction; protein structure function; protozoal genetics; structural biology; three dimensional imaging /topography

DESCRIPTION (provided by applicant): Microtubule-based motors, dyneins, are nano-meter scale protein machineries and convert chemical energy derived from ATP hydrolysis to mechanical movement. The mechanical movement is employed for various essential intracellular motilities, such as axonal transport, chromosome segregation, and flagellar motility. The importance of dynein functions is underlined by several dynein-related diseases, such as Kartagener's syndrome and amyotrophic lateral sclerosis (ALS). However, little is known about how dynein mutations cause the primary cilia diskinesia or even how dyneins convert chemical energy to mechanical movement. Our long-term goal is to understand the structural basis of energy conversion by dynein and its regulation. To this end we will characterize the structure of dynein-microtubule complexes using cryo-electron microscopy and three-dimensional reconstruction. We have recently obtained two initial structures of dynein-microtubule complexes. One is a low resolution structure of the outer arm dynein-microtubule complex, and another is a medium resolution structure of the microtubule binding domain of dynein, the dynein stalk. Our immediate goals are to refine the structure of the outer arm dynein-microtubule complex to 25Angstroms resolution, and to extend the structure of the dynein stalk structure to 10Angstroms resolution. By studying several structures of these complexes in functionally important states, we would like to elucidate conformational changes essential to the energy conversion, motility and regulation of dynein.

描述（由申请人提供）：基于微管的马达（动力蛋白）是纳米级蛋白质机器，可将 ATP 水解产生的化学能转化为机械运动。机械运动用于各种重要的细胞内运动，例如轴突运输、染色体分离和鞭毛运动。几种与动力蛋白相关的疾病，例如卡塔格纳综合征和肌萎缩侧索硬化症（ALS），强调了动力蛋白功能的重要性。然而，人们对动力蛋白突变如何引起初级纤毛运动障碍，甚至动力蛋白如何将化学能转化为机械运动知之甚少。 我们的长期目标是了解动力蛋白能量转换的结构基础及其调控。为此，我们将使用冷冻电子显微镜和三维重建来表征动力蛋白-微管复合物的结构。我们最近获得了动力蛋白-微管复合物的两种初始结构。一个是外臂动力蛋白-微管复合物的低分辨率结构，另一个是动力蛋白微管结合域（动力蛋白茎）的中等分辨率结构。我们的近期目标是将外臂动力蛋白-微管复合物的结构细化至25埃分辨率，并将动力蛋白茎结构的结构延伸至10埃分辨率。通过研究这些复合物在功能上重要的状态下的几种结构，我们希望阐明对于动力蛋白的能量转换、运动和调节至关重要的构象变化。